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Project Name

The ability to manipulate the DNA of an organism is vital to many modern fields of biology. While we have perfected this in common research species such as E. coli, yeast and mouse cells, it is still impossible to transform many other species researchers study. Our project is an attempt to develop transgenics ) techniques for a family of single celled organisms called choanoflagellates. These species are interesting to researchers because they are the closest living relative to our microbial ancestor that became the first multicellular animal. Nicole King, here at UC Berkeley, and other researchers across the globe who study these little creatures are hindered by the inability to genetically manipulate them.

The Berkeley iGEM 2010 team is applying synthetic biology to this problem. We are engineering bacteria that can deliver DNA into the choano. Choanos are predatory, which makes our job a bit simpler. Once our bacteria is engulfed by the choano, it is programmed to burst using a self-lysis device. Proteins we have placed inside the bacteria will then go into action. First, we have designed a vacuole-buster device that will burst the small food membrane holding the bacteria inside the choano, spewing the contents into the cytoplasm of the cell. In the cytoplasm, a transposon/transposase device tagged with a nuclear localization device will move to the nucleus. In the nucleus, the transposase will splice the transposon into the choanoflagellate DNA.

<html> <img src="http://openwetware.org/images/b/b9/Icon_board.png" alt="Resources"> </html>The Team

Teamberk10.jpg Undergrads

Amy Kristofferson
Conor McClune
Tahoura Samad
Christoph Neyer

Graduate Students
Tim Hsiau


Insert names here

Chris Anderson
Terry Johnson
Ladder pic.png

<html> <img src="http://openwetware.org/images/e/e2/Icon_info.png" alt="News" border="0"> </html>Tools

Extremely Important Material

Lab Notebooks


The Details

Gel Pictures
Construction Files
-80 Stocks
Assembly Trees
New Composite Parts
Robot CSV Files


Polymerase Reactions
Wobble Reaction
Cloning by PCR
PCA Gene Synthesis
Colony PCR

Cleanup Reactions
Regular Zymo Cleanup
Small-Frag Zymo Cleanup
Zymo Gel Purification

Digestions Reactions
EcoRI/BamHI Digest of Wobble Products
EcoRI/BamHI Digest of PCR Products
Manual 2ab Assembly Digest
BglII Digest of EIPCR Products
Ligation of EcoRI/BamHI digests
Ligation of EIPCR digests
Analytical digests (Mapping)

Bacteria Manipulations
Transformation by heat-shock
Picking of colonies
Miniprep purification of DNA
Macherey-Nagel Miniprep
8-Strip Machery Nagel Miniprep
Genomic Miniprep
-80 Glycerol Stocks
Making Competent Cells
Small Scale Electrocompetent Prep-Generic

LR Gateway Transfers
EcoRI/BamHI Part Transfers
What do I do with my sequencing data?
Small Scale Competent Cell Transformation
2ab Assembly
Gibson Assembly
Choano Food Preperation

Subgroup Notebooks

<html> <img src="http://openwetware.org/images/b/b9/Icon_board.png" alt="Resources"> </html>Project

Useful Links

Subgroup Strategies, Overview